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Immunochemical determination of DHRS8 and DHRS12 enzymes in human tissues
Svobodová, Markéta ; Novotná, Eva (advisor) ; Zemanová, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Markéta Svobodová Supervisor: RNDr. Eva Novotná, Ph.D Title of diploma thesis: Expression of DHRS8 and DHRS12 enzymes in human tissues Dehydrogenase/reductase (SDR family) member 8 (DHRS8, SDR16C2) and dehydrogenase/reductase (SDR family) member 12 (DHRS12, SDR40C1) are human microsomal enzymes belonging to the superfamily of short-chain dehydrogenases/reductases (SDR). This superfamily represents one of the largest protein groups. SDR enzymes participate in the metabolism of various xenobiotic and endogenous compounds and are involved in physiological and pathological processes. However, there are still many enzymes which are only poorly characterised. To this date, the expression on mRNA level and catalytic activity toward 5α- androstane-3α,17β-diol are the only available information about DHRS8. Moreover, there is still no published information (apart from the prediction) regarding DHRS12. The aim of this study was to examine the protein expression of DHRS8 and DHRS12 enzymes in various human tissues. The tissue samples were collected from four middle aged male subjects after the sudden death without apparent disease. Proteins of interest were detected using western blotting and...
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Preparation of mammalian vectors encoding selected microsomal SDR enzymes
Slezák, Jan ; Hofman, Jakub (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradci Kralove Department of Biochemical Sciences Candidate: Jan Slezák Superviser: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Preparation of mammalian vectors encoding selected microsomal SDR enzymes Microsomal short-chain dehydrogenases/reductases DHRS1, DHRS7 and HSD11b1 (SDR19C1, SDR34C1, SDR26C1) are membrane-bound NAD(P)H-dependent enzymes metabolizing a broad spectrum of carbonyl-bearing substrates. These enzymes from the superfamily of short-chain dehydrogenases/reductases mediate metabolism of endogenous compounds, such as glucocorticoids, retinoids, sfingolipids as well as various xenobiotics. Apart from their biological roles, they participate in the etiology of severe diseases (e.g cancer, Alzheimer disease, obesity etc.). Knowledge of inhibitory and substrate affinity may lead to better understanding of the functions of these enzymes in organism and to the development of new therapeutic approaches. The goal of the present work was to prepare mammalian vectors encoding DHRS1, DHRS7 and HSD11b1. Primer design and cDNA amplification were among the first steps. Primers beared the sequence recognized by restriction endonucleases which was concomitantly present in the multicloning site of the pCI plasmid. Such primer design allowed...
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Constuction of plasmids for human GPX7 gene reporter assay
Hanousková, Barbora ; Matoušková, Petra (advisor) ; Novotná, Eva (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Hanousková Supervisor: Ing. Petra Matoušková, Ph.D Title of diploma thesis: Construction of plasmids for human GPx7 gene reporter assay Glutathione peroxidases (GPx) catalyze the reduction of H2O2 and organic hydroperoxides to H2O or corresponding alcohols. GPx remove compounds dangerous for the cell and protect cells against oxidative stress. Until now, in human genome in total 8 GPx coding genes were nidentified. Thesis deals with GPx7, a monomeric enzyme with antioxidant effect and that participates in protein folding. Lower expression of GPx7 was observed in breast cancer, Barrett's dysplasia and esophageal cancer. GPx7 might be a classic tumor marker and its inactivation can lead to cancer development. Possibly, microRNAs (miRNAs) cause a low expression of the GPx7 gene. MiRNAs are small non-coding single stranded RNA molecules that are significantly involved in regulation of gene expression. Their main function is inhibition of gene expression at post-transcriptional level, thus prevention translation of mRNA into protein. It is caused by specific binding miRNA to binding sites in 3'UTR region of mRNA. The aim of this thesis is the construction of plasmids with ligated...
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Preparation of mammalian vectors encoding selected aldo-keto reductases
Herbolt, Daniel ; Hofman, Jakub (advisor) ; Novotná, Eva (referee)
1 Abstract Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Daniel Herbolt Supervisor: RNDr. Jakub Hofman, Ph.D. Title of diploma thesis: Preparation of mammalian vectors encoding selected aldo-keto reductases Aldo-keto reductases 1B1, 1B10 and 1C1 (AKR1B1, AKR1B10, AKR1C1) are NADPH-dependent enzymes metabolizing a broad spectrum of carbonyl-bearing substrates. These enzymes from the superfamily of aldo-keto reductases mediate the reduction of endogenous compounds such as hormones and sugars as well as various xenobiotics and drugs. Beside their physiological roles, they significantly participate in the genesis of severe diseases (e.g. cancer, diabetes etc.). Inhibition of AKR1B1, AKR1B10 and AKR1C1 is accepted as promising therapeutic approach for the treatment of these diseases. The goal of the present work was to prepare mammalian vectors encoding AKR1B1, AKR1B10 and AKR1C1. Primer design and cDNA amplification were the first steps. Primers contained the sequence recognized by restriction endonucleases present in the multicloning site of the pCI plasmid which allowed for the generation of cohesive ends and the consequent joining of insert with plasmid by ligation. Prepared vectors were transformed into Escherichia coli HB101 bacteria....
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The Acceleration of Communication and Manifestations of Everydayness in the Information Society. Parallel to the Approaches of Paul Virilio and Godfrey Reggio
Novotná, Eva ; Fišerová, Michaela (advisor) ; Sluková, Tereza (referee)
This thesis deals with the introduction of new technologies in the information society, which cause acceleration of aspects of everyday life. The consequence of this situation is the increased level of alienation and anonymity among people in cities. The thesis compares the view of two authors. The first one is a French writer and philosopher Paul Virilio, the other is an American director Godfrey Reggio. Both authors perceive the transformation of today's society negatively and describe the consequences arising from the introduction of new technologies. Each of them uses a different form of expression - Paul Virilio, writing and text, Godfrey Reggio, documentary films. Nevertheless, they have the same opinion on the relevant issues and come to similar conclusions. The aim of this thesis is to compare the two authors and their ideas and propose possible solutions to prevent negative consequences of acceleration of aspects of everyday life. Key words Virilio, Reggio, acceleration, technology, information society, media, communication
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Enzyme assay in clinical laboratory
Macounová, Eva ; Dršata, Jaroslav (advisor) ; Novotná, Eva (referee)
The purpose of this work is to develop a list of commonly used enzymes in clinical biochemistry laboratory . Start of work is dedicated to the importance of enzymes as clinical biochemical analytes. In the second part, I chose eight enzymes and one of isoenzymes , which are mostly determined in clinical laboratories, specifically on my workplace in the private laboratory Prevedig in Prague: lactate dehydrogenase (LD), alanine aminotransferase (ALT), asparátaminotransferase (AST), - glutamyltransferase (GGT), kreatine kinase (CK), creatine kinase, myocardial isoenzyme (CK - MB) , alkaline phosphatase (ALP), amylase (AMS) and lipase (LPS). I focus here on their function in the organism, presence in various tissues and thein importance in the diagnosis of diseases. Then I present the physiological limits in my workplace and the different enzymes assays with the precise emphasis on the methods used in the laboratory Prevedig. In the last chapter is specifically focuses on the importance of enzymes in relation to the setting of acute myocardial infarction. I describe the development of biochemical markers od AIM from the 50s of last century, up to the present day, that is from enzymes, through the gradual discovery of more accurate isoenzymes to the most sensitive markers used at the present. The...
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Prefab house building in Prague in the 1980's: comparison of housing estate Barrandov and Źižkov
Němcová, Alice ; Pech, Milan (advisor) ; Novotná, Eva (referee)
Prefab house building in Prague in the 1980's: comparison of housing estate Barrandov and Žižkov This thesis deals with the prefab house building in Prague in the eighties through the example of Barrandov and Žižkov. It describes further the circumstances of the creation of both settlements, their development plans and final implementation. Taken into account is also an art master plan of both settlements in relation to the general issue of art in public space in the time of normalization. This thesis demonstrates the influence of Jiří Ševčík, Kevin Lynch and Christian Norberg-Schulz on authors of Barrandov housing estate. Basis of these architects are in contrast with the approach of planners of problematical reconstruction of Žižkov. The chapter dedicated to Žižkov describes both unrealized and realized conversion plans, including their development and public acceptance. In conclusion, it compares the two settlements and describes their current status.
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Construction of prefabricated residential houses in the 1970s with emphasis on the construction of "Severní město" in Prague
Kokeš, Václav ; Biegel, Richard (advisor) ; Novotná, Eva (referee)
The present thesis called The Construction of Prefabricated Residential Houses in the 1970s With Emphasis On The Construction of Severní Město in Prague focuses on the specifics of the conception and realization of the residential districts in the northern part of Prague. It describes the political and social background of the period when these districts were emerging and the atmosphere and workflow in the architecture studios in the 70s as well as the relationship with investors and building companies. The next part focuses on the individual quarters of Severní Město - Prosek, Ďáblice and Bohnice. It outlines the historical, geographical and urbanistic context of the place, including the architectural and town planning competitions for these areas. The part dealing with the realization and the current state of these residential districts compares and contrasts the individual quarters, points out their unique characteristics and approaches them in a broader context. The final part details potential critical perspectives on the development of Severní Město and mentions several Czech parallels, such as the residential areas of Juliánov and Lesná in Brno. # Prague, residential quarter, prefabricated house, Severní Město, Prosek, Ďáblice, Bohnice Abstrakt
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